Species A rotavirus NSP3 acquires its translation inhibitory function prior to stable dimer formation
| Autor: | Renato León-Rodríguez, Tzvetanka D. Dinkova, Blanca H. Ruiz-Ordaz, Luis Padilla-Noriega, Ana María Cevallos, Edgar Reyna-Rosas, Hugo I. Contreras-Treviño |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
Rotavirus
0301 basic medicine Polyadenylation viruses Dimer lcsh:Medicine Gene Expression Viral Nonstructural Proteins Virus Replication Pathology and Laboratory Medicine Physical Chemistry Biochemistry chemistry.chemical_compound Chlorocebus aethiops MG132 Medicine and Health Sciences lcsh:Science Multidisciplinary biology Physics Messenger RNA Poxviruses Vaccinia Virus Cell biology Physical sciences Nucleic acids Chemistry Medical Microbiology Viral Pathogens Viruses RNA Viral Pathogens Dimerization Protein Binding Research Article medicine.drug Chemical physics Poly(A)-Binding Proteins Microbiology Rotavirus Infections Cell Line 03 medical and health sciences Virology Genetics medicine Animals Point Mutation Amino Acid Sequence RNA Messenger Microbial Pathogens Adaptor Proteins Signal Transducing Binding Sites 030102 biochemistry & molecular biology Binding protein Point mutation lcsh:R Host Cells Organisms Biology and Life Sciences Proteins Protein Complexes Proteasomes Dimers (Chemical physics) 030104 developmental biology Chemical Properties Proteasome chemistry Protein Biosynthesis Chaperone (protein) Mutation biology.protein Proteasome inhibitor RNA lcsh:Q Protein Translation Protein Multimerization Eukaryotic Initiation Factor-4G DNA viruses Sequence Alignment Viral Transmission and Infection |
| Zdroj: | PLoS ONE PLoS ONE, Vol 12, Iss 7, p e0181871 (2017) |
| ISSN: | 1932-6203 |
| Popis: | Species A rotavirus non-structural protein 3 (NSP3) is a translational regulator that inhibits or, under some conditions, enhances host cell translation. NSP3 binds to the translation initiation factor eIF4G1 and evicts poly-(A) binding protein (PABP) from eIF4G1, thus inhibiting translation of polyadenylated mRNAs, presumably by disrupting the effect of PABP bound to their 3'-ends. NSP3 has a long coiled-coil region involved in dimerization that includes a chaperone Hsp90-binding domain (HS90BD). We aimed to study the role in NSP3 dimerization of a segment of the coiled-coil region adjoining the HS90BD. We used a vaccinia virus system to express NSP3 with point mutations in conserved amino acids in the coiled-coil region and determined the effects of these mutations on translation by metabolic labeling of proteins as well as on accumulation of stable NSP3 dimers by non-dissociating Western blot, a method that separates stable NSP3 dimers from the monomer/dimerization intermediate forms of the protein. Four of five mutations reduced the total yield of NSP3 and the formation of stable dimers (W170A, K171E, R173E and R187E:K191E), whereas one mutation had the opposite effects (Y192A). Treatment with the proteasome inhibitor MG132 revealed that stable NSP3 dimers and monomers/dimerization intermediates are susceptible to proteasome degradation. Surprisingly, mutants severely impaired in the formation of stable dimers were still able to inhibit host cell translation, suggesting that NSP3 dimerization intermediates are functional. Our results demonstrate that rotavirus NSP3 acquires its function prior to stable dimer formation and remain as a proteasome target throughout dimerization. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|